Detection of SF-MDR provirus in chimeric NOD/SCID mice into which MDR1-transduced human CD34+ PBPC were transplanted
| Transduction Group . | NOD/ SCID Set No* . | Human CD45 Cells in BM (%) . | No. of MDR1-PCR+ Mice with SF-MDR/No. Mice Assessed . | |||
|---|---|---|---|---|---|---|
| BM . | Spleen . | BM MNC . | BM PMN . | |||
| Transwell | ||||||
| IL-3 | 3, 4 | 2 ± 1 | NA | NA | 3/4 | 3/4 |
| IL-3 | 5, 7 | 33 ± 11 | 5/5 | 2/2 | 2/2 | 2/2 |
| IL-3 + SCM | 1, 7 | 30 ± 12 | 3/3 | 3/3 | 4/4 | 3/4 |
| IL-3/FL | 3, 4 | 12 ± 6 | NA | NA | 4/4 | 3/4 |
| IL-3/FL | 5, 6, 7, 8 | 15 ± 5 | 12/12 | 4/8 | 8/8 | 4/8 |
| IL-3/FL + SCM | 1, 2, 7 | 23 ± 4 | 1/1 | 0/1 | 3/4 | 2/2 |
| IL-3/IL-6/SCF/FL | 8 | 12 ± 1 | 2/2 | 2/2 | 2/2 | 1/2 |
| FL/T/SCF | 8 | 8 ± 3 | 3/3 | 3/3 | 3/3 | 3/3 |
| FL/T/V | 7 | 1 ± 0.3 | 3/3 | 1/3 | NA | NA |
| Cell-free viral supernatant | ||||||
| IL-3/IL-6/SCF/FL | 10 | 7 ± 1 | 2/2 | NA | 2/2 | 2/2 |
| FL/T/SCF | 10 | 10 ± 3 | 3/3 | NA | 3/3 | 3/3 |
| FL/T/SCF | 9 | 33 ± 7 | 4/4 | NA | NA | NA |
| Transduction Group . | NOD/ SCID Set No* . | Human CD45 Cells in BM (%) . | No. of MDR1-PCR+ Mice with SF-MDR/No. Mice Assessed . | |||
|---|---|---|---|---|---|---|
| BM . | Spleen . | BM MNC . | BM PMN . | |||
| Transwell | ||||||
| IL-3 | 3, 4 | 2 ± 1 | NA | NA | 3/4 | 3/4 |
| IL-3 | 5, 7 | 33 ± 11 | 5/5 | 2/2 | 2/2 | 2/2 |
| IL-3 + SCM | 1, 7 | 30 ± 12 | 3/3 | 3/3 | 4/4 | 3/4 |
| IL-3/FL | 3, 4 | 12 ± 6 | NA | NA | 4/4 | 3/4 |
| IL-3/FL | 5, 6, 7, 8 | 15 ± 5 | 12/12 | 4/8 | 8/8 | 4/8 |
| IL-3/FL + SCM | 1, 2, 7 | 23 ± 4 | 1/1 | 0/1 | 3/4 | 2/2 |
| IL-3/IL-6/SCF/FL | 8 | 12 ± 1 | 2/2 | 2/2 | 2/2 | 1/2 |
| FL/T/SCF | 8 | 8 ± 3 | 3/3 | 3/3 | 3/3 | 3/3 |
| FL/T/V | 7 | 1 ± 0.3 | 3/3 | 1/3 | NA | NA |
| Cell-free viral supernatant | ||||||
| IL-3/IL-6/SCF/FL | 10 | 7 ± 1 | 2/2 | NA | 2/2 | 2/2 |
| FL/T/SCF | 10 | 10 ± 3 | 3/3 | NA | 3/3 | 3/3 |
| FL/T/SCF | 9 | 33 ± 7 | 4/4 | NA | NA | NA |
Bone marrow (BM) and spleen cells were recovered from chimeric NOD/SCID mice 5 to 13 weeks after transplantation. The proportion of human CD45+ cells was determined by fluorescence-activated cell separation. Qualitative PCR for the presence of the SF-MDR provirus was done on either genomic DNA purified from BM and spleen or on mononuclear cells (MNC) or polymorphonuclear (PMN) cells isolated by Ficoll density-gradient separation of whole BM. In all experiments, BM cells or genomic DNA from NOD/SCID mice not given transplanted cells and from control mice that had mock-transduced CD34+ PBPC transplantation was concurrently analyzed to verify the specificity of the PCR reactions for the provirus.
Plus-minus values are mean ± SE.
CD34+ PBPC transplanted per mouse: 1 × 106 for set 3 and 4; 2 × 106 for sets 1, 2, 5, 6, 7, 8, and 10; and 4 × 106 for set 9.